Classifications

• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/50—Pyridazines; Hydrogenated pyridazines
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  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
  • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
  • C07D239/24—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members
  • C07D239/28—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having three or more double bonds between ring members or between ring members and non-ring members with hetero atoms or with carbon atoms having three bonds to hetero atoms with at the most one bond to halogen, directly attached to ring carbon atoms
  • C07D239/30—Halogen atoms or nitro radicals
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
  • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
  • A61K31/00—Medicinal preparations containing organic active ingredients
  • A61K31/33—Heterocyclic compounds
  • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
  • A61K31/54—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with at least one nitrogen and one sulfur as the ring hetero atoms, e.g. sulthiame
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/02—Drugs for disorders of the urinary system of urine or of the urinary tract, e.g. urine acidifiers
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P13/00—Drugs for disorders of the urinary system
  • A61P13/08—Drugs for disorders of the urinary system of the prostate
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P15/00—Drugs for genital or sexual disorders; Contraceptives
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P25/00—Drugs for disorders of the nervous system
  • A61P25/04—Centrally acting analgesics, e.g. opioids
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P3/00—Drugs for disorders of the metabolism
  • A61P3/06—Antihyperlipidemics
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• • A—HUMAN NECESSITIES
  • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
  • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
  • A61P9/00—Drugs for disorders of the cardiovascular system
  • A61P9/06—Antiarrhythmics
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D231/00—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings
  • C07D231/02—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings
  • C07D231/10—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members
  • C07D231/12—Heterocyclic compounds containing 1,2-diazole or hydrogenated 1,2-diazole rings not condensed with other rings having two or three double bonds between ring members or between ring members and non-ring members with only hydrogen atoms, hydrocarbon or substituted hydrocarbon radicals, directly attached to ring carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D233/00—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings
  • C07D233/54—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members
  • C07D233/56—Heterocyclic compounds containing 1,3-diazole or hydrogenated 1,3-diazole rings, not condensed with other rings having two double bonds between ring members or between ring members and non-ring members with only hydrogen atoms or radicals containing only hydrogen and carbon atoms, attached to ring carbon atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D239/00—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings
  • C07D239/02—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings
  • C07D239/06—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member
  • C07D239/08—Heterocyclic compounds containing 1,3-diazine or hydrogenated 1,3-diazine rings not condensed with other rings having one double bond between ring members or between a ring member and a non-ring member with hetero atoms directly attached in position 2
  • C07D239/10—Oxygen or sulfur atoms
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D249/00—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms
  • C07D249/02—Heterocyclic compounds containing five-membered rings having three nitrogen atoms as the only ring hetero atoms not condensed with other rings
  • C07D249/08—1,2,4-Triazoles; Hydrogenated 1,2,4-triazoles
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings
  • C07D401/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
  • C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D403/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00
  • C07D403/02—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings
  • C07D403/12—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, not provided for by group C07D401/00 containing two hetero rings linked by a chain containing hetero atoms as chain links
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D405/00—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom
  • C07D405/14—Heterocyclic compounds containing both one or more hetero rings having oxygen atoms as the only ring hetero atoms, and one or more rings having nitrogen as the only ring hetero atom containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D413/00—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms
  • C07D413/14—Heterocyclic compounds containing two or more hetero rings, at least one ring having nitrogen and oxygen atoms as the only ring hetero atoms containing three or more hetero rings
• • C—CHEMISTRY; METALLURGY
  • C07—ORGANIC CHEMISTRY
  • C07D—HETEROCYCLIC COMPOUNDS
  • C07D491/00—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00
  • C07D491/02—Heterocyclic compounds containing in the condensed ring system both one or more rings having oxygen atoms as the only ring hetero atoms and one or more rings having nitrogen atoms as the only ring hetero atoms, not provided for by groups C07D451/00 - C07D459/00, C07D463/00, C07D477/00 or C07D489/00 in which the condensed system contains two hetero rings
  • C07D491/04—Ortho-condensed systems

Definitions

• ⁇ 1A is the appellation recently approved by the IUPHAR Nomenclature Committee for the previously designated “ ⁇ 1C " cloned subtype as outlined in the 1995 Receptor and Ion Channel Nomenclature Supplement (Watson and Girdlestone, 1995).
• ⁇ 1c is used throughout this application and the supporting tables and figures to refer to the receptor subtype recently renamed " ⁇ 1A ". Since in both the old and new nomenclature there has only been one unique receptor subtype which has been designated ⁇ 1C (i.e., there is no ⁇ 1C under the current nomenclature), " ⁇ 1C " is an unambiguous description of this unique receptor subtype.
• Benign Prostatic Hyperplasia also called Benign Prostatic Hypertrophy
• Benign Prostatic Hypertrophy is a progressive condition which is characterized by a nodular enlargement of prostatic tissue resulting in obstruction of the urethra. This results in increased frequency of urination, nocturia, a poor urine stream and hesitancy or delay in starting the urine flow.
• Chronic consequences of BPH can include hypertrophy of bladder smooth muscle, a decompensated bladder and an increased incidence of urinary tract infection.
• the specific biochemical, histological and pharmacological properties of the prostate adenoma leading to the bladder outlet obstruction are not yet known. However, the development of BPH is considered to be an inescapable phenomenon for the aging male population.
• BPH is observed in approximately 70% of males over the age of 70.
• the method of choice for treating BPH is surgery (Lepor, H., Urol. Clinics North Amer., 17, 651 (1990)). Over 400,000 prostatectomies are performed annually (data from 1986). A medicinal alternative to surgery is clearly very desirable.
• the limitations of surgery for treating BPH include the morbidity rate of an operative procedure in elderly men, persistence or recurrence of obstructive and irritative symptoms, as well as the significant cost of surgery.
• ⁇ -Adrenergic receptors McGrath, et. al. Med. Res.
• ⁇ -Adrenergic drugs can be broken down into two distinct classes: agonists (clonidine and naphazoline are agonists), which mimic the receptor activation properties of the endogenous neurotransmitter norepinephrine, and antagonists (phenoxybenzamine and prazosin are antagonists), which act to block the effects of norepinephrine. Many of these drugs are effective but also produce unwanted side effects (for example, clonidine produces dry mouth and sedation in addition to its antihypertensive effects).
• agonists clonidine and naphazoline are agonists
• phenoxybenzamine and prazosin are antagonists
• ⁇ -adrenergic drugs are not selective for any particular ⁇ -adrenergic receptor. Many of these drugs produce untoward side effects which may be attributed to their poor ⁇ -adrenergic receptor selectivity.
• nonselective ⁇ -antagonists have been prescribed to treat BPH.
• M. Caine, et al. reported that the nonselective ⁇ -antagonist phenoxybenzamine was useful in relieving the symptoms of BPH. This drug may produce its effects by interacting with ⁇ -receptors located on the prostate.
• the ⁇ -adrenergic antagonists prazosin and terazosin have also been found to be useful for treating BPH.
• these drugs also produce untoward side effects.
• the ⁇ 1C receptor is responsible for mediating the contraction of human prostate smooth muscle (Gluchowski, C. et. al., WO 94/10989, 1994; Forray, C. et. al., Mol. Pharmacol. 45, 703, 1994).
• the ⁇ 1C antagonists may be effective agents for the treatment of BPH with decreased side effects.
• the ⁇ 1C receptor may also be present in other lower urinary tract tissues, such as urethral smooth muscle (Ford et al. Br. J. Pharmacol., 114, 24P, (1995)).
• This invention is directed to dihydropyrimidine compounds which are selective antagonists for cloned human ⁇ 1C receptors. This invention is also related to uses of these compounds for lowering intraocular pressure (Zhan, et. al. Ophthalmol. Vis. Sci., 34 Abst.
• This invention is directed to dihydropyrimidine compounds which are selective antagonists for human ⁇ 1C receptors.
• This invention is also related to uses of these compounds for lowering intraocular pressure, inhibiting cholesterol synthesis, relaxing lower urinary tract tissue, the treatment of benign prostatic hyperplasia, impotency, cardiac arrhythmia and for the treatment of any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention further provides a pharmaceutical composition comprising a therapeutically effective amount of the above-defined compounds and a pharmaceutically acceptable carrier.
• the present invention is directed to compounds having the structures :
• each of Y 1 , Y 2 , Y 3 , Y 4 and Y 5 is independently -H; straight chained or branched C 1 -C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; -F, -Cl, -Br, or -I; -NO 2 ; -N 3 ; -CN; -OR 3 , -OCOR 3 , -COR 3 , -CONHR 3 , -CON(R 3 ) 2 , or -COOR 3 ; or any two of Y 1 , Y 2 , Y 3 , Y 4 and Y 5 present on adjacent carbon atoms can constitute a methylenedioxy
• R 2 is -H; straight chained or branched C 1 -C 7 alkyl, hydroxyalkyl, alkoxyalkyl, aminoalkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; C 3 -C 10 cycloalkyl-C 1 -C 10 -alkyl, C 3 -C 10 cycloalkyl-C 1 -C 10 -monofluoroalkyl or C 3 -C 10 cycloalkyl-C 1 -C 10 -polyfluoroalkyl; -CN;
• each p is independently an integer from 1 to 7; where each n is independently an integer from 0 to 5; where each R 3 is independently -H; straight chained or branched C 1 -C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; where R 4 is
• presently preferred compounds include the following:
• the compounds may have the structures:
• V is selected from CR 5 R 7 or NR 7 and p is selected from 1-3.
• the invention further provides that the compound has the following structures :
• the invention further provides that the compound has the structure:
• the invention further provides that the compound has the structures:
• presently preferred compounds include the following:
• presently preferred compounds include the following:
• R 5 is selected from -CO 2 CH 3 or -H.
• the present invention is directed to compounds having the structures:
• each of Y 1 , Y 2 , Y 3 , Y 4 and Y 5 is independently -H; straight chained or branched C 1 -C 7 alkyl, monofluoroalkyl or polyfluoroalkyl; straight chained or branched C 2 -C 7 alkenyl or alkynyl; C 3 -C 7 cycloalkyl, monofluorocycloalkyl, polyfluorocycloalkyl or cycloalkenyl; -F, -Cl, -Br, or -I; -NO 2 ; -N 3 ; -CN; -OR 4 , -OCOR 4 , -COR 4 , -CONHR 4 , -CON(R 4 ) 2 , or -COOR 4 ; or any two of Y 1 , Y 2 , Y 3 , Y 4 and Y 5 present on adjacent carbon atoms can constitute a methylenedi
• the invention further provides for the (-) and (+) enantiomers of the compounds described above.
• (-) and (+) enantiomers of the compounds described above are provided.
• presently preferred compounds include the following:
• presently preferred compounds include the following:
• the invention further provides for a pharmaceutical composition comprising a therapeutically effective amount of any of the compounds described herein and a pharmaceutically acceptable carrier.
• the therapeutically effective amount is an amount from about 0.01 mg per subject per day to about 500 mg per subject per day, preferably from about 0.1 mg per subject per day to about 60 mg per subject per day and most preferably from about 1 mg per subject per day to about 20 mg per subject per day.
• the therapeutically effective amount is an amount from about 0.01 mg to about 500 mg.
• the pharmaceutical carrier may be a liquid and the pharmaceutical composition would be in the form of a solution.
• the pharmaceutically acceptable carrier is a solid and the composition is in the form of a powder or tablet.
• the pharmaceutical carrier is a gel and the composition is in the form of a suppository or cream.
• the compound of the pharmaceutical composition additionally does not cause a fall in blood pressure at dosages effective to alleviate benign prostatic hyperplasia. In a further embodiment the compound of the pharmaceutical composition additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram per rat.
• the invention provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject one of the compounds described herein effective to treat benign prostatic hyperplasia.
• the invention further provides that the compound additionally does not cause a fall in blood pressure at dosages effective to alleviate benign prostatic hyperplasia.
• the compound additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram of rat.
• the compound effects treatment of benign prostatic hyperplasia by relaxing lower urinary tract tissue and in particular where lower urinary tract tissue is urethral smooth muscle.
• the invention further provides a method of treating a subject suffering from high intraocular pressure which comprises administering to the subject one of the compounds described herein effective to lower intraocular pressure.
• the invention further provides a method of treating a subject suffering from a disorder associated with high cholesterol which comprises administering to the subject one of the compounds described herein effective to inhibit cholesterol synthesis.
• the invention also provides a method of treating a disease which is susceptible to treatment by antagonism of the ⁇ 1C receptor which comprises administering to the subject one of the compounds described herein effective to treat the disease.
• the invention further provides a method of treating a subject suffering from impotency which comprises administering to the subject one of the compounds described herein effective to treat impotency.
• the invention further provides a method of treating a subject suffering from sympathetically mediated pain which comprises administering to the subject one of the compounds described herein effective to treat sympathetically mediated pain.
• the invention provides a method of treating a subject suffering from cardiac arrhythmia which comprises administering to the subject one of the compounds described herein effective to treat cardiac arrhythmia.
• the invention provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject one of the compounds described herein in combination with a 5 alpha-reductase inhibitor effective to treat benign prostatic hyperplasia.
• the 5-alpha reductase inhibitor is finasteride.
• a pharmaceutical composition comprising a therapeutically effective amount one of the compounds described herein in combination with a therapeutically effective amount of finasteride and a pharmaceutically acceptable carrier.
• the therapeutically effective amount of one of the compounds described herein is an amount from about 0.01 mg to about 500 mg and the therapeutically effective amount of the finasteride is about 5 mg.
• the therapeutically effective amount one of the compounds described herein is an amount from about 0.1 mg to about 60 mg and the therapeutically effective amount of finasteride is about 5 mg. In a further embodiment of the invention the therapeutically effective amount of the one of the compounds described herein is an amount from about 1 mg to about 20 mg and the therapeutically effective amount of finasteride is about 5 mg.
• the invention further provides a method of relaxing lower urinary tract tissue which comprises contacting the lower urinary tract tissue with an amount of one of the compounds described herein effective to relax lower urinary tract tissue. In one embodiment the lower urinary tract tissue is urethral smooth muscle.
• the invention provides a method of relaxing lower urinary tract tissue in a subject which comprises administering to the subject an amount of one of the compounds described herein effective to relax lower urinary tract tissue.
• the lower urinary tract tissue is urethral smooth muscle.
• the invention provides for the use of the compounds described herein for the preparation of a pharmaceutical composition for lowering intraocular pressure, inhibiting cholesterol synthesis, and the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention provides for the use of the compounds described herein for the preparation of a pharmaceutical composition for relaxing lower urinary tract tissue and in particular urethral smooth muscle.
• the invention further provides for the use of any of compounds described herein for the preparation of a pharmaceutical composition, where the compound additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention provides that the compound used in the preparation of the pharmaceutical composition additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram per rat.
• the invention provides for the use of the compounds described herein in the preparation of a medicament for lowering intraocular pressure, inhibiting cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention provides for the use of the compounds described herein in the preparation of a medicament for relaxing lower urinary tract tissue and in particular urethral smooth muscle.
• the invention further provides for the use of any of compounds described herein in the preparation of a medicament, where the compound additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention further provides that the compound in the medicament additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram per rat.
• the invention provides for a drug which is useful for lowering intraocular pressure, inhibiting cholesterol synthesis, and the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful, the effective ingredient of the said drug being any of the compounds described herein.
• the invention further provides the drug described herein additionally does not cause a fall in blood pressure at dosages effective to lower intraocular pressure, to inhibit cholesterol synthesis, and for the treatment of: benign prostatic hyperplasia, impotency, cardiac arrhythmia and any disease where antagonism of the ⁇ 1C receptor may be useful.
• the invention further provides that the drug additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram per rat.
• the invention provides for a drug which is useful for relaxing lower urinary tract tissue and in particular urethral smooth muscle, the effective ingredient of the drug being any of the compounds described herein.
• the invention further provides the drug which is useful for relaxing lower urinary tract tissue additionally does not cause a fall in blood pressure at dosages effective to relax lower urinary tract tissue.
• the invention further provides that the drug which is useful for relaxing lower urinary tract tissue additionally does not cause a fall in blood pressure in rats at a dosage of 10 micrograms of compound per kilogram per rat.
• the invention also provides for the (-) and (+) enantiomers of all compounds of the subject application described herein. Included in this invention are pharmaceutically acceptable salts and complexes of all of the compounds described herein.
• the salts include but are not limited to the following acids and bases.
• the following inorganic acids hydrochloric acid, hydrofluoric acid, hydrobromic acid, hydroiodic acid, sulfuric acid and boric acid.
• the organic acids acetic acid, trifluoroacetic acid, formic acid, oxalic acid, malonic acid, succinic acid, fumaric acid, tartaric acid, maleic acid, citric acid, methanesulfonic acid, trifluoromethanesulfonic acid, benzoic acid, glycolic acid, lactic acid and mandelic acid.
• the following inorganic bases ammonia, hydroxyethylamine and hydrazine.
• organic bases methylamine, ethylamine, propylamine, dimethylamine, diethylamine, trimethylamine, triethylamine, ethylenediamine, hydroxyethylamine, morpholine, piperazine and guanidine.
• This invention further provides for the hydrates and polymorphs of all of the compounds described herein.
• the invention also provides a pharmaceutical composition comprising a therapeutically effective amount of the compounds described above and a pharmaceutically acceptable carrier.
• a "therapeutically effective amount” is any amount of a compound which, when administered to a subject suffering from a disease against which the compounds are effective, causes reduction, remission, or regression of the disease.
• the therapeutically effective amount is an amount from about 0.01 mg per subject per day to about 500 mg per subject per day, preferably from about 0.1 mg per subject per day to about 60 mg per subject per day and most preferably from about 1 mg per subject per day to about 20 mg per subject per day.
• the "pharmaceutically acceptable carrier” is any physiological carrier known to those of ordinary skill in the art useful in formulating pharmaceutical compositions.
• the invention also provides for pharmaceutical composition
• pharmaceutical composition comprising a therapeutically effective amount of the any of the compounds described herein in combination with a therapeutically effective amount of finasteride and a pharmaceutically acceptable carrier.
• the pharmaceutical composition is a therapeutically effective amount from about 0.01 mg per subject per day to about 500 mg per subject per day of any one of the compounds described herein and a therapeutically effective amount of the finasteride of about 5 mg per subject per day.
• a more preferred embodiment of the pharmaceutical composition is a therapeutically effective amount from about 0.1 mg per subject per day to about 60 mg per subject per day of any one of the compounds described herein and a therapeutically effective amount of the finasteride of about 5 mg per subject per day.
• the most preferred embodiment of the pharmaceutical composition is a therapeutically effective amount from about 1 mg per subject per day to about 20 mg per subject per day of any one of the compounds described herein and a therapeutically effective amount of the finasteride of about 5 mg per subject per day.
• the pharmaceutical carrier may be a liquid and the pharmaceutical composition would be in the form of a solution.
• the pharmaceutically acceptable carrier is a solid and the composition is in the form of a powder or tablet.
• the pharmaceutical carrier is a gel and the composition is in the form of a suppository or cream.
• the compound may be formulated as a part of a pharmaceutically acceptable transdermal patch.
• a solid carrier can include one or more substances which may also act as flavoring agents, lubricants, solubilizers, suspending agents, fillers, glidants, compression aids, binders or tablet-disintegrating agents; it can also be an encapsulating material.
• the carrier is a finely divided solid which is in admixture with the finely divided active ingredient.
• the active ingredient is mixed with a carrier having the necessary compression properties in suitable proportions and compacted in the shape and size desired. The powders and tablets preferably contain up to 99% of the active ingredient.
• Suitable solid carriers include, for example, calcium phosphate, magnesium stearate, talc, sugars, lactose, dextrin, starch, gelatin, cellulose, polyvinylpyrrolidine, low melting waxes and ion exchange resins.
• Liquid carriers are used in preparing solutions, suspensions, emulsions, syrups, elixirs and pressurized compositions.
• the active ingredient can be dissolved or suspended in a pharmaceutically acceptable liquid carrier such as water, an organic solvent, a mixture of both or pharmaceutically acceptable oils or fats.
• the liquid carrier can contain other suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmoregulators.
• suitable pharmaceutical additives such as solubilizers, emulsifiers, buffers, preservatives, sweeteners, flavoring agents, suspending agents, thickening agents, colors, viscosity regulators, stabilizers or osmoregulators.
• suitable examples of liquid carriers for oral and parenteral administration include water (partially containing additives as above, e.g. cellulose derivatives, preferably sodium carboxymethyl cellulose solution), alcohols (including monohydric alcohols and polyhydric alcohols, e.g. glycols) and their derivatives, and oils (e.g. fractionated coconut oil and arachis oil).
• the carrier can also be an oily ester such as ethyl oleate and isopropyl myristate.
• Sterile liquid carriers are useful in sterile liquid form compositions for parenteral administration.
• the liquid carrier for pressurized compositions can be halogenated hydrocarbon or other pharmaceutically acceptable propellent.
• Liquid pharmaceutical compositions which are sterile solutions or suspensions can be utilized by for example, intramuscular, intrathecal, epidural, intraperitoneal or subcutaneous injection. Sterile solutions can also be administered intravenously.
• the compounds may be prepared as a sterile solid composition which may be dissolved or suspended at the time of administration using sterile water, saline, or other appropriate sterile injectable medium.
• Carriers are intended to include necessary and inert binders, suspending agents, lubricants, flavorants, sweeteners, preservatives, dyes, and coatings.
• the compound can be administered orally in the form of a sterile solution or suspension containing other solutes or suspending agents, for example, enough saline or glucose to make the solution isotonic, bile salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitol and its anhydrides copolymerized with ethylene oxide) and the like.
• a sterile solution or suspension containing other solutes or suspending agents, for example, enough saline or glucose to make the solution isotonic, bile salts, acacia, gelatin, sorbitan monoleate, polysorbate 80 (oleate esters of sorbitol and its anhydrides copolymerized with ethylene oxide) and the like.
• compositions suitable for oral administration include solid forms, such as pills, capsules, granules, tablets, and powders, and liquid forms, such as solutions, syrups, elixirs, and suspensions.
• forms useful for parenteral administration include sterile solutions, emulsions, and suspensions.
• Optimal dosages to be administered may be determined by those skilled in the art, and will vary with the particular compound in use, the strength of the preparation, the mode of administration, and the advancement of the disease condition. Additional factors depending on the particular subject being treated will result in a need to adjust dosages, including subject age, weight, gender, diet, and time of administration.
• the invention further provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject an amount of the one the compounds described above effective to treat benign prostatic hyperplasia.
• the invention also provides a method of treating a subject suffering from high intraocular pressure which comprises administering to the subject an amount of any of the compounds described above effective to lower intraocular pressure.
• This invention also provides a method of treating a subject suffering a disorder associated with high cholesterol which comprises administering to the subject an amount of any of the compounds described above effective to inhibit cholesterol synthesis.
• This invention also provides a method of treating a disease which is susceptible to treatment by antagonism of the ⁇ 1c receptor which comprises administering to the subject an amount of any the compounds described above effective to treat the disease.
• This invention also provides a method of treating a subject suffering from impotency which comprises administering to the subject an amount of any of the compounds described above effective to treat impotency.
• This invention also provides a method of treating a subject suffering from sympathetically mediated pain which comprises administering to the subject an amount of any of the compounds described above effective to treat sympathetically mediated pain.
• This invention also provides a method of treating a subject suffering from cardiac arrhythmia which comprises administering to the subject an amount of any of the compounds described above effective to treat cardiac arrhythmia.
• This invention also provides a method of treating a subject suffering from benign prostatic hyperplasia which comprises administering to the subject an amount of any of the compounds described above in combination with a 5 alpha-reductase inhibitor effective to treat benign prostatic hyperplasia.
• the 5-alpha reductase inhibitor is finasteride.
• the dosage administered to the subject is about 0.01 mg per subject day to 50 mg per subject per day of finasteride in combination with an ⁇ 1C antagonist.
• a preferred dosage administered to the subject is about 0.2 mg per subject per day to 10 mg per subject per day of finasteride in combination with an ⁇ 1C antagonist.
• a more preferred dosage administered to the subject is about 1 mg per subject per day to 7 mg per subject per day of finasteride in combination with an ⁇ 1C antagonist.
• the most preferred dosage administered to the subject is about 5 mg per subject per day of finasteride in combination with an ⁇ 1C antagonist.
• the mixture was refluxed for 4.5 hours and then cooled to room temperature.
• Aqueous HCl (6 N, 50 mL) was added and stirring was continued for 1 hour.
• the mixture was basified to pH 9 by addition of 6 N aq.
• This compound was prepared from 1,6-dihydro-5-methoxycarbonyl-2-[ ⁇ (4-methoxyphenyl)methyl ⁇ thio]-4-methyl-6-(4-nitrophenyl)-1-[(4-nitrophenyloxy)carbonyl]pyrimidine (0.592 g, 1 mmol), 3-[4-cyano-4-phenyl
• This compound was prepared from 1,6-dihydro-3- ⁇ N-[3-(4- phenylpiperidin-1-yl)propyl] ⁇ carboxamido-6-methoxycar bonyl-2-[ ⁇ (4-methoxyphenyl)methyl ⁇ thio]-6-(4-nitrophe nyl)-4-methylpyrimidine (0.15 g, 0.223 mmol) using the procedure described in Example 5 and purified by flash column chromatography (0.102 g, 83%); m.p.
• This compound was prepared from 1- ⁇ N-[3-(4-cyano-4-phenylpiperidin-1-yl)propyl] ⁇ carboxamido-1,6-dihydro-6-methoxycarbonyl-2-[ ⁇ (4-methoxyphenyl)methyl ⁇ thio]-6-(4-nitrophenyl)-4-methylpyrimidine (0.15 g, 0.215 mmol) using the procedure described in Example 5 and purified by flash column chromatography (0.118 g, 95%); m.p.
• This compound was prepared from 1,6-dihydro-6-methoxycarbonyl-3- ⁇ N-[4-methoxycarbonyl-phenyl-piperidin-1-yl]propyl ⁇ carboxamido-2-[ ⁇ (4-methoxyphenyl)methyl ⁇ thio]- 6-(4-nitrophenyl)-4-methylpyrimidine (0.730 g, 1 mmol) using the procedure described in Example 5 and purified by flash column chromatography (0.57 g, 94%); m.p.
• pyrimidine 4-(4-Methoxyphenyl)-4-phenylpiperidine. 4-Hydroxy-4-phenylpiperidine (5.00 g, 28.2 mmol) was added to a suspension of AlCl 3 (18.8 g, 0.141 mol, 5.00 eq) in anhydrous anisole (100 mL). The mixture was stirred at room temperature for 1 hours and then heated to 50 °C for 3.5 hours. It was cooled to room temperature and poured cautiously into ice-water. The mixture was basified to pH 11 by addition of 6 N aqueous NaOH, and extracted with EtOAc (3 ⁇ 75 mL).
• This compound was prepared from 1,6-dihydro-5-methoxycarbonyl-2-[ ⁇ (4-methoxyphenyl)methyl ⁇ thio]-4-methyl-6-(4-nitrophenyl)-1-[(4-nitrophenyloxy)carbonyl]pyrimidine (0.592 g, 1 mmol), K 2 CO 3 (0.276 g, 2 mmol), 3-[4-ethoxycarbonyl-4-phenylpiperidin-1-yl]propylamine (0.350 g, 1.2 mmol, 1.2 eq), ethanethiol (0.5 mL), and TFA (0.5 mL) using the procedure described in Example 10 and purified by flash column chromatography (0.295 g, 47%) ; m.p.
• the first major product to elute was (-)-1,6-dihydro-2-methoxy-5-methoxycarbonyl-4-methyl-6-(4-nitrophenyl)-1- ⁇ N-[(2-phenyl)ethyl] ⁇ carboxamidopyrimidine and this compound was crystallized from isopropyl ether (0.85 g, 33.6%); m.p.
• phenyl)-2-oxo-1,2,3,6-tetrahydropyrimidine-5-carboxylic acid (0.2 g, 0.346 mmol), 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.382 g, 2 mmol), and 4-(N,N-dimethylamino)pyridine (0.488 g, 4 mmol), in methanol (20 mL) was stirred and refluxed for 5 h and the solvent evaporated. The residue was redissolved in CH 2 Cl 2 (15 mL), washed with saturated aqueous ammonium chloride solution (3 X 10 mL), and dried (Na 2 SO 4 ).
• (+)-5-(2-cyanoethoxycarbonyl)-4-ethyl-1,6-dihydro-2-methoxy-6- (4-nitrophenyl)-1- ⁇ N-[(2-phenyl)ethyl] ⁇ carboxamido pyrimidine (2.62 g, 5.182 mmol) in toluene (40 mL) was added 1,8-diazabicyclo[5,4,0]-undec-7-ene (0.237,1.55 mmol) at room temperature and the resulting solution was heated at 90 °C for 3.5 minutes.
• Scheme 5 a. Benzyl 3-[(3,4-difluorophenyl)methylene]-4-oxopentanoate.
• Second major product to elute was (-)-5-(benzyloxycarbonyl)-4-ethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6-(3,4-diflurophenyl)pyrimidine. Colorless oil.
• (+)-5-(Benzyloxycarbonyl)-1,6-dihydro-2-methoxy-4-ethyl-6-(3,4-diflurophenyl)pyrimidine To a stirred solution of (+)-5-(benzyloxycarbonyl)-4-ethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6- (3,4-diflurophenyl)pyrimidine (1.83 mmol, 1.0 g) in toluene (10 mL) was added 1,8-diazabicyclo[5,4,0]-undec-7-ene (0.81 mmol, 0.12 mL) at room temperature and the resulting solution was heated to reflux for 5h and then stirred for 12h at room temperature.
• (+)-5-(benzyloxycarbonyl)-1,6-dihydro-2-methoxy-4-ethyl-6- (3,4-diflurophenyl)pyrimidine (17.0 g, 44.04 mmol) and 4-dimethylaminopyridine (6.99 g, 57.25 mmol) in CH 2 Cl 2 (200 mL) was added a powder of 4-nitrophenyl chloroformate 11.54 g, 57.25 mmol) at room temperature. The reaction mixture was stirred for 12 hours and then the solvent was removed in vacuo.
• (+)-6-(3,4-difluorophenyl)-4-ethyl-1- ⁇ N-[3-(4-methoxycarbonyl-4-phenylpiperidin-1-yl)propyl] ⁇ carboxamido-1,2,3,6-tetrhydro-2-oxopyrimidine-5-carboxylic acid (0.22 g, 0.375 mmol) in CH 2 Cl 2 (3 mL) was added 4-N,N-dimethylamino pyridine (0.14 g, 1.12 mmol) and 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (0.21 g, 1.12 mmol) under argon and the resulting solution was stirred at room temperature for 2h.
• 1-Benzyl-4-carboxamido-4-(2-pyridyl)piperidine To 1-benzyl-4-cyano-4-(2-pyridyl)piperidine (4.5 g, 14.3 mmol), 10 ml of conc. H 2 SO 4 was added and the solution was stirred at room temperature for 24 hours. It was cooled to 0 °C, diluted with ice pieces and poured into crushed ice. The mixture was then carefully neutralized with 50 % NaOH solution.
• 5-Methylbenzfuroxan 4-Methyl-2-nitroaniline (100 g, 0.650 mol) was suspended in saturated alcoholic sodium hydroxide solution (1.50 1). To this suspension was added with cooling (5 °C) commercial aqueous sodium hypochlorite till the red color disappeared. The fluffy yellow precipitate formed was filtered, washed with cold water and recrystallized from ethanol to get 5-Methylbenzfuroxan (88.2 g, 89 % yield) as a pale solid.
• 5-Methylbenzofurazan To 5-Methylbenzfuroxan (88.2 g, 0.59. mol) in refluxing EtOH (75 ml) was added dropwise P(OEt) 3 (150 ml ). When addition was complete, refluxing was continued for 1 more hour. The solvent was removed by rotary evaporation and the residue shaken with water (200 mL) and allowed to stand overnight at (0-5 °C). The brown solid so obtained was filtered, washed with water and chromatograghed on silica gel to yield 5-Methylbenzofurazan (70 g, 87 %) as white needle.
• 5-Dibromomethylbenzofurazan 5-Methylbenzofurazan (70 g, 0.52 mol), NBS (325 g), and benzoyl peroxide (0.5 g) were refluxed with stirring in carbon tetrachloride (1.5 L) with exclusion of moisture for 30 hours. The reaction mixture was washed with water (2X0.5L), dried (NaSO 4 ), and the solvent was removed in vacuo. The residue was chromatographed (silica, EtOAc- hexane, 1:150) to give 122 g (80%) of the title compound. The resulting white solid was used in the next step without any further characterization.
• Examples 26 and 27 are two diastereomeric products derived from the (+) enantiomer at the pyrimidine part and the two possible enantiomeric compounds with respect to the fluoromethylene chiral center.
• Example 28 is two diastereomeric products derived from the (+) enantiomer at the pyrimidine part and the two possible enantiomeric compounds with respect to the fluoromethylene chiral center.
• the resulting mixture of diastereomers was separated by column chromatography (SiO 2 , 3% EtOAc in toluene).
• the first major product to elute was (+)-5-(benzyloxycarbonyl)-4-ethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6-(2,4-difluorophenyl)pyrimidine (12.15 g, 38%).
• the second major product to elute was the other diastereomer and no effort was made to isolate it.
• (+)-5- (benzyloxycarbonyl)-1,6-dihydro-2-methoxy-4-ethyl-6- (2,4-difluorophenyl)pyrimidine as a viscous oil (6.15 g, 78%).
• (+)-5-(2-cyanoethoxycarbonyl)-4-methoxymethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6-(3,4-difluorophenyl)pyrimidine (2.80 g, 5.46 mmol) in toluene (80 mL) was added 1,8-diazabicyclo[5,4,0]-undec-7-ene (0.250 g, 1.64 mmol) and the mixture was stirred at 75 °C for 1 h.
• (+)-5-(2-cyanoethoxycarbonyl)-1,6-dihydro-2-methoxy-4- methoxymethyl-6-(3,4-difluorophenyl)pyrimidine as a viscous oil (0.82 g, 40.5%).
• the first major product to elute was (+)-5-methoxycarbonyl-4-methoxmethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6- (3,4-difluorophenyl)pyrimidine (1.74 g, 44.5%).
• the second major product to elute was the other diastereomer and no effort was made to isolate it.
• (+)-5-methoxycarbonyl-4-methoxymethyl-1,6-dihydro-1- ⁇ N-[2-phenyl)ethyl] ⁇ carboxamido-2-methoxy-6-(3,4-difluorophenyl)pyrimidine (1.74 g, 3.67 mmol) in toluene (40 mL) was added 1,8-diazabicyclo[5,4,0]-undec-7-ene (0.250 g, 1.64 mmol) and the mixture was stirred at 70-80 °C for 1.5 h.
• (+)-5-methoxycarbonyl- 4-methoxymethyl-1,6-dihydro-2-methoxy-6-(3,4-difluorophenyl)-1-[(4-nitrophenyloxy)carbonyl]pyrimidine (1.30 g, 78%) as a viscous oil.
• the HCl salt was prepared by treatment with 1N HCl in ether; m.p.
• (+)-6-(3,4-Difluorophenyl)-1,6-dihydro-2-oxo-5-methoxy- carbonyl-4-bromomethyl-1- [(4-nitrophenyloxy)carbonyl]pyrimidine (1.5 mmol, 0.81 g) was heated in oil bath for 3 h (bath temperature 130°C).
• (+)-6-(3,4-difluorophenyl)-1,2,3,6- tetrahydro-2-oxo-5-methoxycarbonyl-4-methyl-1-(3- bromopropylaminocarbonyl)pyrimidine (0.435 g, 1.0 mmol) and 1- (2-carboxamidophenyl)piperazine (0.4 g, 2.0 mmol) in 25 mL of anhydrous acetone was added powdered K 2 CO 3 (0.69 g, 5.0 mmol) and KI (0.17 g, 1.0 mmol) and the resulting suspension was heated to reflux for 10 h.
• (+)-1,2,3,6-tetrahydro-1- ⁇ N-[4-(2-carboxamidophenyl)piperazin-1yl]propyl ⁇ -carboxamido-4-methyl-6- (3,4-difluorophenyl)-2-oxo-pyrimidine was obtained as light yellow powder (0.48 g, 84% yield).
• Benzyl 3-oxo-2-(2,4-difluorobenzylidenyl)butanoate A mixture of 2, 4-difluorobenzaldehyde (7.1 g, 50 mmol.), benzyl acetoacetate (12.48 g, 65 mmol.), acetic acid (0.15 g, 2.5 mmol.), piperidine (0.212 g, 2.5 mmol.) and 2-propanol (300 mL) was stirred at room temperature for two days. After the removal of solvent, the residue was then dissolved in ethyl acetate and washed with saturated KHSO 4 , saturated NaHCO 3 , water and then dried over Na 2 SO 4 .
• an oral composition of a compound of this invention 100mg of one of the compounds described herein is formulated with sufficient finely divided lactose to provide a total amount of 580 to 590 mg to fill a size O hard gel capsule.
• Binding affinities were measured for selected compounds of the invention at six cloned human alpha-1 and alpha-2 receptor subtypes, as well as at the L-type calcium channel. The protocols for these experiments are given below. Protocol for the Determination of the Potency of ⁇ 1 Antagonists
• ⁇ 1A Human Adrenerglc Receptor The entire coding region of ⁇ 1A (1719 bp), including 150 base pairs of 5' untranslated sequence (5' UT) and 300 bp of 3' untranslated sequence (3' UT), was cloned into the BamHI and ClaI sites of the polylinker-modified eukaryotic expression vector pCEXV-3, called EXJ.HR.
• the construct involved the ligation of partial overlapping human lymphocyte genomic and hippocampal cDNA clones: 5' sequence were contained on a 1.2 kb SmaI-XhoI genomic fragment (the vector-derived BamHI site was used for subcloning instead of the internal insert-derived SmaI site) and 3' sequences were contained on an 1.3 kb XhoI-ClaI cDNA fragment (the ClaI site was from the vector polylinker).
• Stable cell lines were obtained by cotransfection with the plasmid ⁇ 1A/EXJ (expression vector containing the ⁇ 1A receptor gene) and the plasmid pGCcos3neo (plasmid containing the aminoglycoside transferase gene) into LM(tk-), CHO, and NIH3T3 cells, using calcium phosphate technique.
• the cells were grown, in a controlled environment
• ⁇ 1B ' Human Adrenerglc Receptor The entire coding region of ⁇ lB (1563 bp), including 200 base pairs and 5' untranslated sequence (5' UT) and 600 bp of 3' untranslated sequence (3' UT), was cloned into the EcoRI site of pCEXV-3 eukaryotic expression vector. The construct involved ligating the full-length containing EcoRI brainstem cDNA fragment from ⁇ ZapII into the expression vector. Stable cell lines were selected as described above.
• ⁇ 1C Human Adrenerglc Receptor The entire coding region of ⁇ 1C (1401 bp), including 400 base pairs of 5' untranslated sequence (5' UT) and 200 bp of 3' untranslated sequence (3' UT), was cloned into the KpnI site of the polylinker-modified pCEXV-3-derived eukaryotic expression vector, EXJ.RH.
• the construct involved ligating three partial overlapping fragments: a 5' 0.6kb HincII genomic clone, a central 1.8 EcoRI hippocampal cDNA clone, and a 3' 0.6Kb PstI genomic clone.
• the hippocampal cDNA fragment overlaps with the 5' and 3' genomic clones so that the HincII and Pstl sites at the 5' and 3' ends of the cDNA clone, respectively, were utilized for ligation.
• This full-length clone was cloned into the KpnI site of the expression vector, using the 5' and 3' KpnI sites of the fragment, derived from vector (i.e., pBluescript) and 3'-untranslated sequences, respectively.
• Stable cell lines were selected as described above.
• Radioligand Binding Assays Transfected cells from culture flasks were scraped into 5ml of 5mM Tris-HCl, 5mM EDTA, pH 7.5, and lysed by sonication. The cell lysates were centrifuged at 1000 rpm for 5 min at 4°C, and the supernatant was centrifuged at 30,000 ⁇ g for 20 min at 4°C. The pellet was suspended in 50mM Tris-HCl, 1mM MgCl 2 , and 0.1% ascorbic acid at pH 7.5.
• Binding of the ⁇ 1 antagonist [ 3 H] prazosin (0.5 nM, specific activity 76.2 Ci/mmol) to membrane preparations of LM(tk-) cells was done in a final volume of 0.25 ml and incubated at 37°C for 20 min. Nonspecific binding was determined in the presence of 10 ⁇ M phentolamine. The reaction was stopped by filtration through GF/B filters using a cell harvester. Inhibition experiments, routinely consisting of 7 concentrations of the tested compounds, were analyzed using a non-linear regression curve-fitting computer program to obtain Ki values.
• ⁇ 2 Human Adrenergic Receptors To determine the potency of ⁇ 1 antagonists at the ⁇ 2 receptors, LM(tk-) cell lines stably transfected with the genes encoding the ⁇ 2A , ⁇ 2B , and ⁇ 2C receptors were used.
• the cell line expressing the ⁇ 2A receptor is designated L- ⁇ 2A , and was deposited on November 6, 1992 under ATCC Accession No. CRL 11180.
• the cell line expressing the ⁇ 2B receptor is designated L-NGC- ⁇ 2B , and was deposited on October 25, 1989 under ATCC Accession No. CRL10275.
• the cell line expressing the ⁇ 2C receptor is designated L- ⁇ 2C , and was deposited on November 6, 1992 under ATCC Accession No. CRL-11181.
• Cell lysates were prepared as described above (see Radioligand Binding Assays), and suspended in 25mM glycylglycine buffer (pH 7.6 at room temperature). Equilibrium competition binding assay were performed using [3H] rauwolscine (0.5nM), and nonspecific binding was determined by incubation with 10 ⁇ M phentolamine. The bound radioligand was separated by filtration through GF/B filters using a cell harvester.
• the compounds described above were assayed using cloned human alpha adrenergic receptors and the rat calcium channel.
• the preferred compounds were found to be selective ⁇ 1C antagonists.
• the binding affinities of compounds 19-23 are illustrated in the following table. Binding affinities of compounds 19-23 at cloned human ⁇ 1a, ⁇ 1b and ⁇ lc receptors.

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